小鼠白细胞介素18基因克隆、测序及重组质粒pIRES-IL18-B7.1的构建

Cloning and Sequencing of Mouse IL-18 and Constructionof Recombinant Plasmid pIRES-IL18-B7.1

目的 :克隆小鼠白细胞介素 1 8( m IL- 1 8)编码区的 c DNA序列并构建其表达载体。方法 :利用逆转录多聚酶链反应 ( RT- PCR)法 ,以小鼠脾细胞 m RNA为模板 ,扩增获得全长 m IL-1 8,与 p MD1 8T连接作全自动测序 ,并利用基因重组技术构建包含 m IL- 1 8和 B7.1的双基因表达质粒 p IRES- IL1 8- B7.1。结果 :经测序证实获得的 m IL- 1 8序列与文献报道完全一致 ,并成功构建了表达质粒。结论 :成功克隆了 m IL- 1 8的 c DNA,并构建了双基因表达载体 p IRES- IL1 8- B7.

Objective: To clone the sequence of the cDNA of mouse IL 18 coding area and construct its expression vector. Methods: Using mouse splenocyte mRNA as template to obtain full length mIL 18 with the technique of RT PCR followed by automatic sequencing of pMD18T mIL 18 and to construct a recombinant plasmid containing mIL 18 and B7.1 genes with recombinant DNA technique. Results: Sequencing proved the cloned mIL 18 cDNA to be completely identical with that reported in the literature and the recombinant plasmid containing mIL 18 and B7.1 was constructed successfully. Conclusion: mIL 18 cDNA was successfully cloned and an expression vector pIRES IL18 B7.1 was constructed.